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Updated: Aug 9, 2026
Whole-animal Imaging and Flow Cytometric Techniques for Analysis of Antigen-specific CD8+ T Cell Responses after Nanoparticle Vaccination
Published on: April 29, 2015
Detection of antigen-specific T cell cytokine expression in whole blood by flow cytometry
M A Suni1, L J Picker, V C Maino
1Becton Dickinson Immunocytometry Systems, San Jose, CA 95131, USA. maria_suni@bdis.com
Insights
A simplified flow cytometry method rapidly detects antigen-specific T cells in whole blood, offering insights into immune responses and drug effects.
Area of Science:
- Immunology
- Cell Biology
- Flow Cytometry
Background:
- Flow cytometry enables detection of single cell cytokine expression for T cell characterization.
- Previous methods required peripheral blood mononuclear cell (PBMC) cultures.
Purpose of the Study:
- To describe a simplified procedural modification for detecting low-frequency memory T cells in whole blood.
- To compare T cell responses in whole blood versus PBMC cultures.
- To assess the utility of this method for clinical applications.
Main Methods:
- A modified flow cytometry technique was developed for whole blood analysis.
- Detection of cytokine-expressing CD4+ and CD8+ T cells in response to soluble antigen.
- Comparison of T cell responses between whole blood and PBMC cultures.
- Investigation of T cell responses to cytomegalovirus (CMV) in seropositive individuals.
- Blocking experiments using anti-class II major histocompatibility complex (MHC) antibodies.
Main Results:
- The simplified method rapidly detects low-frequency memory CD4+ and CD8+ T cells expressing cytokines in whole blood.
- Whole blood cultures showed similar, slightly higher percentages of antigen-responsive T cells compared to PBMC cultures.
- Cytomegalovirus-specific T cell responses were detected in whole blood only from seropositive individuals.
- CD4+ T cell responses were inhibited by anti-class II MHC antibodies.
Conclusions:
- This simplified whole blood flow cytometry procedure is effective for detecting antigen-specific memory T cells.
- The method allows for the examination of T cell immunity in clinical samples.
- It may be useful for assessing the direct effects of pharmacological drug concentrations on T cell immunity.
Abstract:
We have recently described a highly sensitive flow cytometric technique, based on the ability to detect single cell expression of cytokines, to simultaneously quantitate and phenotypically characterize antigen-specific memory/effector T cells in PBMC cultures. In this report, we describe a simplified procedural modification which enables the rapid detection of low frequency memory CD4+ and CD8+ T cells expressing cytokines in response to soluble antigen in whole blood. When compared with T cell responses in PBMC cultures, whole blood cultures demonstrated similar but slightly higher percentages of T cells responsive to specific antigen. In addition, T cell responses to cytomegalovirus in whole blood were observed only in sensitized (seropositive) individuals, and CD4+ T cell responses could be blocked by anti-class II MHC antibodies. This procedure may provide a means to examine direct effects of pharmacological drug concentrations on T cell immunity in clinical samples.
